Product Highlight

• It is produced on the B+A grade cGMP sterile canned production line, with components completely limited-- the number of protein types doesn’t exceed 20 and the total content is less than 1.5 mg/mL, all of which are artificially recombinant, without human-derived or animal-derived components. It has been filed with the US FDA DMF filing No. 37873.
• Primary cell separation and continuous subculturing to P20 can be achieved without changing the culture medium.
• The products can improve cell adhesion and adaptability, hence suitable for large-scale production of cell products and 3D culture systems, and more suitable for constructing cell banks used to support clinical applications.

Product Information

This culture medium can be used for the primary cell isolation, seed - bank construction, and continuous subculture of human mesenchymal stem cells from various sources, while maintaining their multi – lineage differentiation potential, such as bone marrow (BM - hMSC) and umbilical cord (UCM - hMSC). No serum or serum substitutes are required when using this product.

This product has a well-defined chemical composition, with no added serum or serum substitutes (platelet lysate), and no components of animal or human origin. There is little batch - to - batch variation. All raw materials meet GMP standards, making it more suitable for clinical research purposes.

Product Specifications:

Osmotic pressure	280-350mOsm/kg
Endotoxin	<0.25EU/mL
Sterility	Bacteria and fungi shall not be detected
Mycoplasma	shall not be detected
pH	6.8-7.8
Appearance	Light yellow - yellow clear and transparent liquid
Volume	500mL/bottle

Special Instructions Before Use

1. The usage method of this product is indeed different from that of serum - containing culture media and serum - substitute culture media.

We kindly request users to carefully read the instruction manual of this product before operation. Please do not use this product solely based on experience.

Compared with serum - containing culture media and serum - substitute culture media, certain details of the existing operations have been standardized. (For example, there are clear requirements for the operation method of adding the culture medium to the culture flask or petri dish.)

2. This product does not contain a trypsin inhibitor and cannot be used to terminate trypsin digestion. Serum and serum substitutes (platelet lysate) contain antithrombin III, which can inhibit the digestive effect of trypsin. This is why serum can terminate trypsin digestion. It is also the reason why MSC cells in serum - containing culture media must be washed with PBS before digestion (to remove residual serum) during sub -culture.

When using this product, it is recommended to use GMP - grade stem cell mild digestive enzyme (a supporting product, product number: LNC1004.1) to digest cells. This method does not require an inhibitor for termination. After dilution with the culture supernatant, the cells can be collected by centrifugation to remove the supernatant. Moreover, it causes no damage to the cells within 10 minutes, and the harvested cells have a high viability. The multiple amplification remains at a relatively high level after sub - culture, avoiding the problem of trypsin residue after cell digestion.

Data from Continuous subculturing

Passage	Initial cells	cell culture type	Number of tissues
P0	4.50E+05	60mm dishes	4
		Increase by folds	Seeding density (cells/cm2)
P1	2.50E+06	10.23	10000
P2	2.28E+06	9.11
P3	2.49E+06	9.97
P4	2.00E+06	8.01
P5	2.01E+06	8.03
P6	2.35E+06	7.82	12000
P7	2.17E+06	5.79
P8	1.64E+06	5.45
P9	1.59E+06	5.3
P10	1.58E+06	5.25
P11	1.91E+06	5.1	15000
P12	1.52E+06	4.04
P13	1.25E+06	3.34
P14	1.41E+06	3.77
P15	1.06E+06	2.83

The inoculum size is provided below if petri dish of different surface area is used:

Culture flask/dish	Approximate bottom area (cm2)	Day 0 medium volume (mL)	Day 2 medium volume (mL)	Tissue segment added (number)
60 mm dish	21	2	3	5
100mm dish	55	5	7	12
T-25 flask	25	2	3	6
T-75 flask	75	7	8	15
T-175 flask	175	15	20	40
T-225 flask	225	20	25	50